CN107567447A - The dimer impurity and its minimizing technology of Eliquis - Google Patents

The dimer impurity and its minimizing technology of Eliquis Download PDF

Info

Publication number
CN107567447A
CN107567447A CN201680026473.6A CN201680026473A CN107567447A CN 107567447 A CN107567447 A CN 107567447A CN 201680026473 A CN201680026473 A CN 201680026473A CN 107567447 A CN107567447 A CN 107567447A
Authority
CN
China
Prior art keywords
formula
eliquis
dimer impurity
impurity
compound
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201680026473.6A
Other languages
Chinese (zh)
Inventor
埃莱娜·布拉索拉
菲利波·托马西
洛里斯·佩鲁齐
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fabbrica Italiana Sintetici SpA (FIS)
Original Assignee
Fabbrica Italiana Sintetici SpA (FIS)
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fabbrica Italiana Sintetici SpA (FIS) filed Critical Fabbrica Italiana Sintetici SpA (FIS)
Publication of CN107567447A publication Critical patent/CN107567447A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D11/00—Solvent extraction
    • B01D11/02—Solvent extraction of solids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B63/00—Purification; Separation; Stabilisation; Use of additives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/28—Control of physical parameters of the fluid carrier
    • G01N30/36—Control of physical parameters of the fluid carrier in high pressure liquid systems
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • G01N30/7233—Mass spectrometers interfaced to liquid or supercritical fluid chromatograph
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N2030/022—Column chromatography characterised by the kind of separation mechanism
    • G01N2030/027—Liquid chromatography

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Animal Behavior & Ethology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Veterinary Medicine (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Analytical Chemistry (AREA)
  • Hematology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Diabetes (AREA)
  • Urology & Nephrology (AREA)
  • Vascular Medicine (AREA)
  • Cardiology (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
  • Hydrogenated Pyridines (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

The purpose of the present invention is active component Eliquis (Apixaban) dimer impurity, the analysis method for identifying and/or quantifying it and the synthetic method for removing the impurity in the impurity or limitation Eliquis and its synthesis precursor from Eliquis and its synthesis precursor.

Description

The dimer impurity and its minimizing technology of Eliquis
Technical field
The present invention relates to active component Eliquis (Apixaban) impurity, the impurity can be in synthesis Eliquis Period produces；Method for identifying and/or quantifying these impurity；With for Eliquis and its synthesis precursor in remove institute State synthetic method existing for impurity or the limitation impurity.
The invention further relates to purposes of these dimer impurities in analysis method.
Background technology
Eliquis is a kind of active pharmaceutical ingredient for the anti-coagulants for being used as treatment venous thrombo-embolic event.In addition, It also shows the prospect for the treatment of acute coronary syndrome (ACS), cerebrovascular ischemia and cancer.
Eliquis shown in below formula (I)
With chemical name 1- (4- methoxyphenyls) -7- oxos -6- [4- (2- oxo-piperidine -1- bases) phenyl] -4,5- two Hydrogen pyrazolo [5,4-c] pyridine-3-carboxamide and CAS RN 503612-47-3.
Have been described above several methods for preparing Eliquis.
Specifically, WO2007/001385 discloses a kind of synthetic method of more kilogramme-scales.
Specifically, WO2007/001385 disclosed in embodiment 6 it is a kind of according to following reaction process to 10Kg scales Ah Piperazine sand class ethyl ester carries out amidation process to prepare the method for Eliquis：
According to described program, reacted at least 12 hours, obtained using the anhydrous ammonia in propane diols and at 90 DEG C The molar yield that must be separated is 94.6% Eliquis.
Jian'an Jing and YafeiJi Synthetic Communications, 43,72-79,2013 disclose one kind The cost-effective strategy of Eliquis is synthesized, is summarized in described disclose in the flow 3 of page 74.
The method that another kind prepares Eliquis is disclosed in WO 03/049681；Particularly in embodiment 54 to 56, Which depict the route of synthesis that the active pharmaceutical ingredient is prepared since compound 10 (intermediate of Eliquis).Specifically Ground, in embodiment 54, in part B final section, 3- is produced by means of Ammonia and EtOAc mixture Quinoline -4- bases -1- [4- (2- oxo piperidine -1- bases)-phenyl] -5,6- dihydro -1H- pyridin-2-ones (63)；Finally by fast Compound 63 is obtained after speed is column chromatography eluting.But by means of filtering and then sequentially using in the part A of the same embodiment Ethanol and ether are washed to prepare different compounds, are referred to as copper (I), a kind of catalyst.Gained copper (I) catalytic reaction is to produce Raw compounds 63, therefore washed without above-mentioned filtering and sequentially with ethanol and ether to produce compound 63, but only need Remove copper removal (I).
Another method for obtaining Eliquis intermediate is disclosed in CN104030972.Especially disclose in embodiment 1 The method for obtaining compound (I) (i.e. the intermediate of Eliquis).Specifically, in step (3), the compound is by means of adding Water is added then to filter acquisition；Yield is finally produced as 74.0% and compound (I) of the purity higher than 98.5%.
In laboratory experiment, surprisingly it has been found that a variety of route of synthesis for being used to prepare Eliquis disclosed in document carry For the product polluted by HMW impurity.
The presence of the macromolecule impurity, the amount for being especially higher than 0.10% is present, in the medicine of such as Eliquis It is unacceptable in active material.
The content of the invention
Therefore, the present invention solves the problems, such as to be to provide a kind of improved method for being used to prepare Eliquis, methods described Allow to prepare the Eliquis of the HMW impurity with low content.
This problem is addressed by the method for preparing Eliquis described in claims, the claim The definition of book is the part of this specification.
Specifically, in the first aspect, the present invention provide one kind prepare substantially free of following structure (II) and/or (III) method of the active component Eliquis of dimer impurity：
On the other hand, the present invention provides a kind of dimer impurity of the formula (II) determined in Eliquis and/or formula (III) Method.
On the other hand, the present invention provides the formula (II), (III), (IV), (V), (VI) and (VII) as new chemical entities Compound.
On the other hand, the present invention provides the impurity of the formula (II), (III), (IV), (V), (VI) and (VII) of separation true Determine in the identity of impurity described in the sample or its precursor of Eliquis and/or the method for amount as " reference mark thing " and/or The purposes of " normative reference thing ".
Brief description of the drawings
Fig. 1 shows the main-process stream that Eliquis is synthesized in prior art program, and it includes the chemical constitution and amount with impurity Carriage of the impurity represented Jing Guo the building-up process.
Fig. 2 shows the main-process stream of the synthesis Eliquis according to the inventive method, according to including step a) to method d) Obtain, and the main-process stream includes passing through taking for the building-up process with the impurity that the chemical constitution of impurity and amount represent Band situation.
Fig. 3 shows that the UV of the dimer impurity of the Eliquis of the formula (II) by LC-MS measurements is composed.
Fig. 4 shows that the UV of the dimer impurity of the Eliquis of the formula (III) by LC-MS measurements is composed.
Embodiment
The purpose of the present invention is a kind of method for preparing formula (I) Eliquis：
The Eliquis includes the dimer impurity of the following Eliquis less than 0.10%：
(E) -6,6'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (1- (4- of following structure (II) Methoxyphenyl) -7- oxos -4,5,6,7- tetrahydrochysene -1H- pyrazolos [3,4-c] pyridine-3-carboxamide)：
And/or the dimer impurity of following Eliquis：
Double (4- (3- carbamyls -1- (4- the methoxyphenyls) -7- oxos -4,5- of (Z) -1,2- of following structure (III) Dihydro-1 h-pyrazole simultaneously [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
Wherein it the described method comprises the following steps：
A) morpholinyl -1- (4- (2- oxo-piperidine -1- bases) phenyl) -5,6- dihydropyridines -2 of formula (IX) are prepared Solution of (1H) -one in the mixture of C1-C3 alcohol and water：
B) solution prepared in filtration step a) is to remove following dimer impurity：
(E) -1,1'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) of formula (VI) it is double (morpholinyl -5, 6- dihydropyridines -2 (1H) -one) and/or formula (VII) double (4- (morpholinyl -2- oxo -5, the 6- dihydros of impurity (Z) -1,2- Pyridine -1 (2H)-yl) phenyl) phenodiazine olefinic oxide：
C) solution of formula (IX) compound comprising purifying is obtained in filtrate,
D) step c) formula (IX) compound is converted into formula (I) Eliquis；
Or alternatively, step a) is replaced to d) with following steps：
E) prepare formula (VIII) 1- (4- methoxyphenyls) -7- oxos -6- (4- (2- oxo-piperidine -1- bases) phenyl) - Solution of 4,5,6,7- tetrahydrochysene -1H- pyrazolo [3,4-c] the Nicotinicum Acidum ethyl esters in C1-C4 alcohol：
F) solution prepared in filtration step e) is to remove following dimer impurity：
(E) -6,6'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (1- (4- methoxies of lower formula (IV) Base phenyl) -7- oxo -4,5,6,7- tetrahydrochysene -1H- pyrazolo [3,4-c] Nicotinicum Acidums ethyl ester) and/or formula (V) impurity (Z) double (4- (3- (ethoxy carbonyl) -1- (4- the methoxyphenyls) -7- oxo -4,5- dihydro-1 h-pyrazoles simultaneously [3,4- of -1,2- C] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
G) solution of formula (VIII) compound comprising purifying is obtained in filtrate,
H) optionally, separate formula (VIII) compound of the purifying of solid form, and repeat step e) to g) once or Repeatedly,
I) formula by step g) or h) (VIII) compound is converted into formula (I) Eliquis.
In fact, have surprisingly found that bag filter contains the alcohol solution and/or formula (VIII) chemical combination of formula (IX) compound The alcoholic solution of thing is possible to optionally remove formula (VI) and (VII), formula (IV) dimer impurity related to (V's) respectively.Cause This, the impurity retains on the filter in solid form, while, the purified product of all amounts is collected into the molten of filtration In liquid.Retain the product of solid matter on the filter actually without formula (IX) or (VIII), and only containing formula (VI) and Or the dimer impurity of (IV) and (V) and optional other inorganic filter aids and/or activated carbon (VII).
An advantage of the present invention is to allow that the precursor for preparing following Eliquis and Qi Shi (IX) and (VIII), Its contain less than 0.10% be respectively formula (II) and (III), formula (VI) and (VII), formula (IV) and (V) each dimer it is miscellaneous Matter.
Another epochmaking advantage of the inventive method is that methods described extremely effective reduces the water of dimer impurity It is flat, while extremely effective removing dimer impurity and avoiding spawn from losing, i.e., provide production with almost quantitative yield Thing.This result is better than the result as obtained by any other recrystallization method certainly, in any other recrystallization side In method, although can obtain identical dimer impurity reduction level in theory, the yield for being purified product must be than passing through The almost quantitative yield that the inventive method obtains is much lower.
In addition, as shown in Figure 1, it has been observed that the dimer impurity is very difficult to remove, in fact, it is present in Ah piperazine's sand In each step of class's synthetic method, because during intermediate separates, it can not simultaneously lose, therefore be contained with high dimer impurity Amount is reached in product Eliquis, through all synthesis steps of method.
Finally, it is necessary to consider that because these dimer impurities of Eliquis have HMW, it is in HPLC High relative retention time is shown during analysis, or pole is frequently, for analysis Eliquis and its related impurities (such as isomers Impurity) analysis method in, the dimer impurity is retained by chromatographic column, and in most cases, it can not be eluted, So as to be retained on column cap.Accordingly, it is determined that the presence of the dimer impurity is not it is clear that also, consequently, because measure The presence of dimer impurity is simultaneously non-obvious.It is also such for the situation of the method for removal or the reduction of the impurity.
Lower formula (IX) compound can be according to Synthetic Communication, volume 2013,43, in the 72-79 pages It is prepared by disclosed synthetic method：
The compound of lower formula (VIII) can be prepared according to the method in above-mentioned article：
Eliquis prepared according to the methods of the invention includes the Eliquis of the formula (II) and (III) less than 0.10% Each in dimer impurity, wherein 0.10% by analyzing HPLC A/A% measure.The measure can use embodiment 8 Described in analysis method advantageously carry out.
In the step a) of the inventive method, morpholinyl -1- (4- (2- oxo-piperidine -1- bases) benzene of formula (IX) is prepared Base) solution of (1H) -one of -5,6- dihydropyridines -2 in the mixture of C1-C3 alcohol and water：
The solution for preparing formula (IX) compound refers to formula (IX) compound of solid form being dissolved in C1-C3 alcohol and water In mixture, or alternatively, (IX) is dissolved in C1-C3 alcohol, then adds water.
Alternatively, the solution for preparing formula (IX) compound can also be by will be another in the mixture of C1-C3 alcohol and water Kind compound (i.e. precursor) is converted into formula (IX) compound to realize.
Alternatively, prepare formula (IX) compound solution can also by C1-C3 alcohol by another compound (before i.e. Body) formula (IX) compound is converted into, water is then added to realize.
Alternatively, prepare formula (IX) compound solution can also by another solvent by another compound (i.e. precursor) is converted into formula (IX) compound, then changes the solvent mixture of C1-C3 alcohol and water into or changes C1-C3 alcohol into Then water is added to realize.
According to a preferred embodiment, another compound (i.e. precursor) can be such as formula (X) compound：
According to a preferred embodiment, formula (X) compound is converted into formula (IX) compound by means of anti-with chlorine valeric chloride Should come carry out.
According to a preferred embodiment, in the mixture of C1-C3 alcohol and water, C1-C3 alcohol is ethanol.
According to a preferred embodiment, solvent is changed to C1-C3 alcohol and then addition water.It is highly preferred that the conversion of solvent is used Ethanol is carried out, and then adds water.
Morpholinyl -1- (4- (2- oxo-piperidine -1- bases) benzene of the formula (IX) prepared in the step a) of the inventive method Base) -5,6- dihydropyridines -2 (1H) -one：
Solution in the mixture of C1-C3 alcohol and water is milky white solution or the fine suspension or outstanding of a small amount of solid matter Supernatant liquid, typically, the amount of the insoluble substance are less than 5 weight % in terms of formula (IX) compound by weight.In fact, described molten In liquid, formula (VI) and (VII) dimer impurity are retained in the form of insoluble solid, hence it is demonstrated that the solution of compound (IX) Or outside the fine suspension of a small amount of solid matter or the milky of suspension being made up of the dimer impurity of formula (VI) and (VII) See.Formula (IX) compound is then dissolved completely in the solution of C1-C3 alcohol and waters.
In the mixture of C1-C3 alcohol and water, C1-C3 alcohol is the unitary selected from methanol, ethanol and normal propyl alcohol or isopropanol Alcoholic solvent.
In the step a) of the inventive method, with formula (IX) compound phase ratio, the amount of C1-C3 alcohol is 2 to 30 volumes.
Volume refers to the volume of per unit product solvent, thus, for example 1 volume be 1 liter every kilogram, or 1 milliliter every gram or 1 microlitre every milligram.Therefore, 10 volumes refer to that for example every kilogram of material (in this case, refers to every kilogram of formula (IX) chemical combination Thing) 10 liters.
According to the inventive method preferred embodiment, in step a), with formula (IX) compound phase ratio, C1-C3 The amount of alcohol is 5 to 20 volumes.
According to the inventive method more preferably embodiment, in step a), with formula (IX) compound phase ratio, C1- The amount of C3 alcohol is 10 to 15 volumes, again it is highly preferred that volume is about 13 volumes.
In the step a) of the inventive method, with formula (IX) compound phase ratio, the amount of water is 0.5 to 20 volume.
According to the inventive method preferred embodiment, in step a), with formula (IX) compound phase ratio, the amount of water For 1 to 15 volume.
According to the inventive method more preferably embodiment, in step a), and formula (IX) compound phase ratio, water Measure as 7 to 13 volumes, it is highly preferred that being about 10 volumes.
According to the inventive method preferred embodiment, in step a), the amount of C1-C3 alcohol is 5 to 20 volumes, and And the amount of water is 1 to 15 volume, both of which and formula (IX) compound phase ratio.
According to the inventive method more preferably embodiment, in step a), the amount of C1-C3 alcohol is 10 to 15 bodies Product, and the amount of water is 7 to 13 volumes, both of which and formula (IX) compound phase ratio, again it is highly preferred that the amount of C1-C3 alcohol is About 13 volumes, and the amount of water is about 10 volumes, both of which and formula (IX) compound phase ratio.
Specifically, in step a), volume ratio between the amount of mixture reclaimed water and the amount of C1-C3 alcohol be included in water with In the mixture of C1-C3 alcohol in the range of 2.4% to 80% (v/v%) water, the mixing preferably in above-mentioned water and C1-C3 alcohol 6.25% to 60% in thing in the range of (v/v%) water, and more preferably in the mixture of above-mentioned water and C1-C3 alcohol 35% to In the range of 50% (v/v%) water.
According to the inventive method preferred embodiment, in step a), C1-C3 alcohol is ethanol.
According to the inventive method more preferably embodiment, in step a), the amount of ethanol is 10 to 15 volumes, and And the amount of water is 7 to 13 volumes, both of which and formula (IX) compound phase ratio, again it is highly preferred that the amount of ethanol is about 13 volumes, And the amount of water is about 10 volumes, both of which and formula (IX) compound phase ratio.
The step a) of the inventive method can be carried out in the temperature between 20 DEG C to 120 DEG C.
According to the inventive method preferred embodiment, temperature of the step a) between 30 DEG C to 50 DEG C is carried out.
According to the inventive method preferred embodiment, temperature of the step a) between 40 DEG C to 45 DEG C is carried out.
Carried out according to the inventive method more preferably embodiment, a temperature of the step a) between 30 DEG C to 50 DEG C, And the amount of ethanol is that the amount of 10 to 15 volumes and water is 7 to 13 volumes, both of which and formula (IX) compound phase ratio.
It is highly preferred that temperature of the step a) between 40 DEG C to 45 DEG C is carried out, and the amount of ethanol is about 13 volumes and water Amount be about 10 volumes, both of which and formula (IX) compound phase ratio.
According to the inventive method preferred embodiment, in step a), filter aid and/or activity are further added Charcoal.
After preparation process a) solution, in step b), the solution is filtered to remove the dimerization of formula (VI) and (VII) Body impurity.
The solution that can be prepared on filter paper or filter paper plate or diatomite plate in filtration step a).
According to the inventive method preferred embodiment, in step b), carried out by means of diatomite plate.
The step b) of the inventive method can be carried out in the temperature between 20 DEG C to 120 DEG C.
According to the inventive method preferred embodiment, temperature of the step b) between 30 DEG C to 50 DEG C is carried out.
According to the inventive method preferred embodiment, temperature of the step b) between 40 DEG C to 45 DEG C is carried out.
According to the inventive method preferred embodiment, in step b), the phase filtered is further used in The C1-C3 alcoholic solvent washing filters of synthermal preheating.
Insoluble formula (VI) and (VII) impurity are retained on the filter in the form of solid matter, and in following feelings Under shape, i.e., filter aid and/or activated carbon are further added wherein in step a) or is carried out in step b) on diatomite plate Filtering, these materials are also contained on filter cake.
In step c), the solution of formula (IX) compound comprising purifying is obtained in filtrate.
In step d), according to known art methods (such as Synthetic Communications, 43；72- The method disclosed in method or preferred WO2007/001385 disclosed in 79,2013) will middle formula (IX) chemical combination obtained of step c) Thing is converted into formula (I) Eliquis.
It is above-mentioned to i) replacing with following steps e) according to the always alternative method based on identical inventive concept of the present invention Step a) is to d).
In step e), 1- (4- the methoxyphenyls) -7- oxos -6- (4- (2- oxo-piperidines -1- of formula (VIII) are prepared Base) phenyl) solution of -4,5,6,7- tetrahydrochysene -1H- pyrazolo [3,4-c] the Nicotinicum Acidum ethyl esters in C1-C4 alcohol：
The solution of the compound (VIII) is prepared in step e) and can be similar to and is changed for the formula (IX) in step a) (including all versions having been described above) described in compound is carried out.
C1-C4 alcohol is selected from by the group of the monohydric alcohol of following material composition：Methanol, ethanol, normal propyl alcohol, isopropanol, n-butanol, Isobutanol, sec-butyl alcohol and the tert-butyl alcohol.
According to the inventive method preferred embodiment, in step e), C1-C4 alcohol is ethanol.
In the step e) of the inventive method, with formula (VIII) compound phase ratio, the amount of C1-C4 alcohol is 3 to 20 volumes.
According to the inventive method preferred embodiment, in step e), with formula (VIII) compound phase ratio, C1- The amount of C4 alcohol is 5 to 15 volumes, more preferably about 10 volumes.
According to the inventive method, temperature of the step e) between 40 DEG C to alcohol of boiling point is carried out.
It is (warm i.e. in backflow in the temperature of the boiling point of alcohol according to the inventive method more preferably embodiment, step e) Degree) carry out.
According to the inventive method preferred embodiment, step e) under the boiling point of ethanol with formula (VIII) chemical combination Thing is carried out compared to the ethanol of 10 volumes.
According to the inventive method preferred embodiment, filter aid and/or activity are further added in step e) Charcoal.
According to the inventive method more preferably embodiment, the further addition activated carbon in step e).
After preparation process e) solution, in step f), the solution is filtered to remove formula (IV) and/or formula (V) Dimer impurity.
The solution that can be prepared on filter paper or filter paper plate or diatomite plate in filtration step e).
The step f) of the inventive method can be carried out in the temperature between 40 DEG C to alcohol of boiling point.
According to the inventive method preferred embodiment, step f) is carried out in the boiling point of alcohol.
According to the inventive method preferred embodiment, in step f), the phase filtered is further used in The C1-C4 alcoholic solvent washing filters of synthermal preheating.
Insoluble formula (IV) and (V) impurity are retained on the filter in the form of solid matter, and in following situation Under, i.e., filter aid and/or activated carbon are further added wherein in step e) or was carried out in step f) on diatomite plate Filter, these materials are also contained on filter cake.
Optionally, formula (IV) and the residual impurity of (V) remain above particularly in formula (VIII) compound of purifying In the case of 0.10%, formula (VIII) compound that can be isolated and purified in solid form in step h) and weight can be passed through Multiple step e) is further purified to g) one or many.
In step i), according to known art methods (such as Synthetic Communications, 43；72- Method disclosed in 79,2013 or WO2007/001385) by step g) or h) in formula (VIII) compound of purifying for obtaining turn Turn to formula (I) Eliquis.
According to the inventive method preferred embodiment, Eliquis includes the following Eliquis less than 0.10% Dimer impurity：Double (4- (3- carbamyls -1- (4- the methoxyphenyls) -7- oxos -4,5- of (Z) -1,2- of lower formula (III) Dihydro-1 h-pyrazole simultaneously [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
The dimer impurity of removal formula (VII) wherein in step b), or in step f) removal formula (V) impurity.
The dimer impurity and its precursor of formula (III) have specific structure, make it more because it carries electron charge Have polarity, so it is less readily soluble in organic solvent, while, it therefore can be more easily in the step b) or f) of the present invention In be removed.
According to the inventive method preferred embodiment, Eliquis include formula (II) less than 0.01% and/or The dimer impurity of formula (III).
According to the inventive method preferred embodiment, Eliquis contains what is determined by HPLC A/A% 0.00% to 0.02% formula (II) and/or the dimer impurity of formula (III), and methods described is included formula (IX) compound Be converted into Eliquis, and wherein described formula (IX) compound include by HPLC A/A% determine 0.1% to 2% it Between the formula (VI) of amount and/or the dimer impurity of formula (VII).
According to the inventive method preferred embodiment, Eliquis contains what is determined by HPLC A/A% 0.02% to 0.10% formula (II) and/or the dimer impurity of formula (III), and methods described is included formula (IX) compound Be converted into Eliquis, and wherein described formula (IX) compound include by HPLC A/A% determine between 2% to 5% The formula (VI) of amount and/or the dimer impurity of formula (VII).
Specifically, from the formula (VI) comprising 2% to 5% amount and/or formula (IX) chemical combination of the dimer impurity of formula (VII) Thing starts, it then follows as discussed previously comprising step a), the inventive method b), c) and d), obtains and contains 0.02% to 0.10% Formula (II) and/or formula (III) dimer impurity Eliquis.
According to the inventive method preferred embodiment, the Eliquis obtained in step d) or step i) has N-1 forms.
Eliquis N-1 forms refer to the Eliquis with the polymorphic Form for being referred to as N-1, the polymorph Form is the solid form for fully being characterized and being prepared, and in EP3009435 embodiment, the form is defined as Ah piperazine The Thermodynamically stable form of husky class.In addition, particularly pass through several technology in the EXPERIMENTAL SECTION EXAMPLES 3 of the application (such as FT-IR, DSC, TGA and X-RPD) characterizes Eliquis form N-1, and its data is incorporated herein by reference.
According to the inventive method preferred embodiment, the described method comprises the following steps a), b), c), so as to obtain The compound (IX) that must be purified, and comprising consecutive steps e), f), g), h), i), therefore obtain formula (I) Eliquis.
In fact, by carry out step a) to c) and carry out step e) to i) the two, it is possible to realized in Eliquis The formula (II) of floor level and/or the impurity of (III).Specifically, the dimer impurity of the formula (II) and/or formula (III) Horizontal further reduce is because all steps of the inventive method are applied in combination.These results are obtained by means of following methods, Since step a), filtered in step b) with removal formula (VI) dimer impurity and/or formula (VII) dimer impurity, Finally obtain the solution of formula (IX) compound comprising purifying.Newest compound is converted into formula (VIII) compound, in step Rapid e) then after step f), filtering formula (VIII) compound is miscellaneous to remove formula (IV) dimer impurity and/or formula (V) dimer Matter, therefore the solution for including formula (VIII) compound purified is obtained in step g), step h) can be optionally carried out, finally Compound (VIII) is converted into formula (I) Eliquis in step i), therefore it contains a small amount of formula (II) dimer impurity And/or formula (III) dimer impurity.
Specifically, what is had been described above is preferably as it allows comprising step a), the inventive method b), c) and d) Dimer impurity is reduced earlier, i.e., the amount of dimer impurity is reduced in the early stage step for prepare the method for Eliquis, from And dimer impurity is avoided carrying around by building-up process and avoids the higher level intermediate with compared with high additive value Loss.
In addition, it is preferably as it more effectively reduces dimerization comprising step a), methods described b), c) and d) Body impurity, particularly formula (VI) dimer impurity and formula (VII) dimer impurity, i.e., its with higher degree from dimer impurity Purifying is provided.
During the inventive method is developed, it has been found that the presence of some impurity, it is very difficult to remove during synthesis, and most Final drug products Eliquis is polluted afterwards.Specifically, pollution formula (IX) and (VIII) are early observed in synthesis The presence of the impurity of precursor intermediate is synthesized, it is the impurity with high relative retention time in HPLC chromatogram.
Therefore, another surprised aspect of the invention is the presence for finding the impurity in Eliquis and its precursor, and And surprisingly it has been found that property, structure and the source of the impurity.
In fact, it has surprisingly been found that turn in formula (XI) compound carried out by means of vulcanized sodium to formula (X) compound During change：
Formula (VI) and (VII) dimer impurity are formed as the accessory substance of the reaction：
In addition, it has surprisingly been found that the impurity advances by Eliquis building-up process, progress identical chemistry turns Change, so as to produce the dimer impurity of the formula of formula (VIII) compound (IV) and (V), then produce the formula of compound Eliquis (II) and (III) dimer impurity.
Fig. 1 depicts the source of impurity and Carriage in Eliquis building-up process.
In addition, it is contemplated that the carrying of impurity, foregoing to be had the following advantages that comprising step a), method b), c) and d)： Dimer impurity is reduced in synthesis early stage, that is, reduces formula (VI) dimer impurity and/or formula into formula (IX) compound (VII) amount of dimer impurity, so as to reduce follow-up intermediate and corresponding related dimer impurity in Eliquis.
Therefore, one aspect of the present invention is Eliquis or the dimer impurity of its precursor, and it is selected from following group：
A (E) -6,6'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (1- (4- first of formula (II) under) Phenyl) -7- oxos -4,5,6,7- tetrahydrochysene -1H- pyrazolos [3,4-c] pyridine-3-carboxamide)：
B double (4- (3- carbamyls -1- (4- the methoxyphenyls) -7- oxos -4,5- two of (Z) -1,2- of formula (III) under) Hydrogen -1H- pyrazolos [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
C (E) -1,1'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (3- morpholines of formula (VI) under) Base -5,6- dihydropyridines -2 (1H) -one)：
D double (4- ((the 2H)-yl of morpholinyl -2- oxo -5,6- dihydropyridines -1) benzene of (Z) -1,2- of formula (VII) under) Base) phenodiazine olefinic oxide：
E (E) -6,6'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (1- (4- first of formula (IV) under) Phenyl) -7- oxo -4,5,6,7- tetrahydrochysene -1H- pyrazolo [3,4-c] Nicotinicum Acidums ethyl ester)：
F double (4- (3- (ethoxy carbonyl) -1- (4- the methoxyphenyls) -7- oxos -4,5- of (Z) -1,2- of formula (V) under) Dihydro-1 h-pyrazole simultaneously [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
The dimer impurity of above-mentioned Eliquis or its precursor is yellow powder.
Specifically, the dimer impurity of the Eliquis obtained by HPLC/DAD detectors is reported in Fig. 3 and Fig. 4 UV-VIS is composed.
Therefore, dimer impurity is the coloured reason of Eliquis.Specifically, the impurity assigns Eliquis with Huang Color.Therefore, it is desirable to Eliquis shows white without the impurity.
On the other hand, it has been found that the dimer impurity of Eliquis and its precursor, particularly formula (II) and (III) Eliquis impurity, especially insoluble in any solvent.
(Z) -1,2- of lower formula (III) it is double (4- (3- carbamyls -1- (4- methoxyphenyls) -7- oxo -4,5- dihydros - 1H- pyrazolos [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
It is the preferred dimer impurity of formula (I) Eliquis.The impurity is yellow powder.
Specifically, there is the dimer impurity of the formula (III) following UV to compose, as shown in figure 4, it is at 360-370nm The higher absorption of display.Therefore, the dimer impurity is the material of display glassy yellow.
The product of chemical reaction is rarely the single compound that purity is sufficient for administrative standard.Agents useful for same in reaction Accessory substance is possibly also present in the product of separation caused by side reaction.Active component (such as Eliquis) production process In some steps, typically by means of high performance liquid chromatography (HPLC), LC/MS, gas chromatography (GC) or thin-layered chromatography (TLC) purity assay, to determine if to be applied to subsequent treatment and finally be applied to drug products.
In general, impurity is identified with spectroscopic methodology, therefore by chromatogram peak position (peak position of such as chromatogram) or TLC Spot on plate is associated with.
After peak position is associated with specific impurities, the impurity in sample can be carried out with regard to its relative position in chromatogram Identification, the wherein position in chromatogram are used in the number of minutes that injection sample and Impurity elution pass through between detector in post and measured.Color The ratio that position in spectrum is known as between retention time and retention time is known as relative retention time.
The technical staff of drug field knows that relatively pure compound may be used as normative reference thing.Normative reference species are seemingly In reference mark thing, but the former can be not only used for checked for impurities, and can be used in the sample of qualitative activity composition being deposited Impurity amount.
As known to the person skilled in the art, the control of Process Impurity is by understanding its chemical constitution, route of synthesis and knowledge Not Ying Xiang in final products the parameter (such as by means of DOE) of the amount of impurity and substantially improve.
The impurity of Eliquis, including do not react completely intermediate, the impurity of raw material, byproduct of reaction, degraded production Thing and other products, the quality and efficiency of the medicament forms containing Eliquis may be influenceed.Therefore, it is necessary to which a kind of determination is miscellaneous The horizontal method of matter, wherein the dimer impurity of Eliquis in Eliquis sample is paid special attention to, particularly a large amount of production medicines During thing Eliquis.
A kind of thus, it is found that dimerization of the Eliquis of lower formula (II) determined in Eliquis and/or lower formula (III) The method of body impurity：
In fact, the dimer impurity of formula (II) and/or formula (III) can use according to analysis below method：In Ah piperazine Sha Banzhong is identified and/or the dimer impurity of quantitative formula (II) and formula (III).
A kind of method for being used to detecting or identifying the dimer impurity of formula (II) and/or formula (III) in Eliquis, its Comprise the following steps：
A) formula (II) of known quantity and/or the dimer impurity of formula (III) are added into Eliquis sample,
B) HPLC or LC/MS analyses are carried out to step a) Eliquis sample,
C) HPLC the or LC/MS peaks of the dimer impurity of formula (II) and/or formula (III) are detected；
Or,
A1 the dimer impurity of formula (II) and/or formula (III)) is analyzed by means of HPLC or LC/MS,
B1 Eliquis sample) is analyzed by means of HPLC or LC/MS,
C1) dimer impurity of formula (II) and/or formula (III) is detected by comparing retention time or relative retention time HPLC or LC/MS peaks；
Or
Detection [M+1] is analyzed by means of LC/MS+Equal to 751 or 767amu peak detects.
In addition to the impurity peaks in Eliquis are identified, according to another aspect of the present invention, it has also been found that a kind of quantitative Ah The method of the dimer impurity of the Eliquis of lower formula (II) and/or lower formula (III) in piperazine sand class：
It comprises the following steps：
A) by means of each list in HPLC or LC/MS measurement Eliquis samples corresponding to formula (II) and/or formula (III) The peak area of one dimer impurity, the Eliquis sample have these compounds of unknown quantity,
B) it is miscellaneous corresponding to the dimer of the formula (II) containing known quantity and/or formula (III) by means of HPLC or LC/MS measurements The peak area of " the normative reference thing " of matter,
C) dimerization in Eliquis is determined with the middle areas measured of step b) by the area of measurement in comparison step a) The amount of body impurity.
According to the inventive method preferred embodiment, for determining the Ah piperazine of the lower formula (III) in Eliquis The method of the dimer impurity of husky class：
Include each foregoing step with the method for formula (III) dimer impurity in quantitative Eliquis, respectively Ground, for the method for determining dimer impurity in Eliquis, comprising a) to c1), and for husky for quantitative Ah piperazine The method of dimer impurity in class, comprising a) to c).
Another aspect of the present invention is the dimer impurity of following Eliquis：
A) formula (II) dimer impurity：
B) formula (III) dimer impurity：
It may be used as " the reference mark for identifying and/or quantifying the dimer impurity in formula (I) Eliquis Thing " or " normative reference thing "：
According to the inventive method preferred embodiment, the dimer impurity of Eliquis is the dimerization of formula (III) Body impurity：
It may be used as " the reference mark for identifying and/or quantifying the dimer impurity in formula (I) Eliquis Thing " or " normative reference thing ".
In addition, following dimer impurity may be used as identifying and/or quantifying described two in following related compound " the reference mark thing " or " normative reference thing " of aggressiveness impurity：
C) in the compound of lower formula (X)：
Formula (IV) dimer impurity and/or formula (V) dimer impurity：
D) in following formula (XI) compound：
Formula (VI) dimer impurity and/or formula (VII) dimer impurity：
According to the inventive method preferred embodiment, by LC/MS perform in quantitative Eliquis 1 To the amount between 100 PPMs lower formula (II) and/or lower formula (III) Eliquis dimer impurity amount side Method：
According to another aspect of the present invention, the dimer impurity of the Eliquis of formula (II) and/or formula (III)：
It can be prepared according to the method comprised the following steps：
A) morpholinyl -1- (4- (2- oxo-piperidine -1- bases) phenyl) -5,6- dihydropyridines -2 of formula (IX) are prepared Solution of (1H) -one in the mixture of C1-C3 alcohol and water：
B) solution prepared in filtration step a),
C) solid on de-entrainment filter, it includes following dimer impurity：Formula (VI) dimer impurity and/or formula (VII) dimer impurity：
D) formula (VI) and the dimer impurity of (VII) are separated,
E) impurity of step d) formula (VI) and/or formula (VII) is converted into formula (II) and/or the Eliquis of (III) Dimer impurity.
Or alternatively, step a) is replaced to e) with following steps：
F) prepare formula (VIII) 1- (4- methoxyphenyls) -7- oxos -6- (4- (2- oxo-piperidine -1- bases) phenyl) - Solution of 4,5,6,7- tetrahydrochysene -1H- pyrazolo [3,4-c] the Nicotinicum Acidum ethyl esters in C1-C4 alcohol：
G) solution prepared in filtration step f),
H) solid on de-entrainment filter, it includes following dimer impurity：Formula (IV) dimer impurity and/or formula (V) Dimer impurity：
I) dimer impurity of formula (IV) and/or formula (V) is separated,
Step i) formula (IV) compound and/or compound (V) j) are converted into Ah's piperazine of formula (II) and/or formula (III) The dimer impurity of husky class.
Separating step d) and i) (such as by preparation HPLC, preparative TLC or tradition can be passed through by means of chromatographic isolation Chromatographic column) carry out.In the case of the latter, can easily use silicone filler tradition chromatographic column, and by using heptane/ AcOEt mixture, preferred gradient elution, separates compound.
The impurity of step d) formula (VI) and/or formula (VII) is converted into formula (II) and/or the Eliquis of (III) The step e) of dimer impurity follows the disclosed phase that formula (IX) compound is converted into formula (VIII) compound in the prior art Same method, its significant difference are to replace parent material, i.e. compound (IX) with the impurity of formula (VI) and/or formula (VII).Specifically Ground, it then follows this method, obtain formula (IV) impurity and/or formula (V) impurity.Then, by the impurity finally obtained (IV) and/or (V) be converted into formula (II) and/or the Eliquis of (III) impurity follow it is disclosed by formula (VIII) compound in the prior art The same procedure for being converted into formula (I) compound is carried out, and its significant difference is to replace starting with the impurity of formula (IV) and/or formula (V) Material, i.e. compound (VIII).
Method of the prior art on preparing Eliquis, including formula (X) compound is prepared by using vulcanization sodium reduction, Such as Synthetic Communications, 43；72-79, the method disclosed in 2013, there is provided containing 0.28% to 1.0% Between amount formula (II) and (III) impurity Eliquis.
Another aspect of the invention is Eliquis itself, because the inventive method, which allows to prepare to contain, passes through HPLC A/ The Eliquis of 0.00% to 0.10% formula (II) of A% measure and/or the dimer impurity of formula (III)；The amount is limited In formula (VI) and/or the initial amount of first impurity of (VII), it produces during formula (X) compound is prepared and is brought into formula (IX) without changing chemical constitution and relative quantity in compound.
Specifically, from the formula (VI) of the amount present in formula (IX) compound between 5% to 2% and/or formula (VII) Dimer impurity starts, it then follows the inventive method, particularly step a) is to d), it is allowed to which the dimer impurity is reduced or gone Remove, reach the formula (II) and/or (III) included in Eliquis by HPLC A/A% 0.10% to 0.02% amounts determined Impurity degree.
Alternatively, from the formula (VI) and/or formula of the amount being included in present in formula (IX) compound between 2% to 0% (VII) dimer impurity starts, it then follows the inventive method, particularly step a) can subtract the dimer impurity to d) Less or remove, reach included in Eliquis by the formulas (II) of HPLC A/A% 0.02% to 0.00% amounts determined and/ Or the degree of the impurity of (III) (referring to Fig. 2).
Therefore, another aspect of the invention is final product Eliquis, determined by HPLC A/A%, it has few The formula (II) of amount and/or the dimer impurity of formula (III).
Therefore, specifically, another aspect of the invention is contain 0.02% to 0.10% determined by HPLC A/A% Formula (II) and/or formula (III) dimer impurity Eliquis, the Eliquis passes through step a) to present invention d) Method obtain, the step is included the formula (VI) comprising the amount between 5% to 2% determined by HPLC A/A% And/or formula (IX) compound of the dimer impurity of formula (VII) is converted into Eliquis.
According on the other hand, determined by HPLC A/A%, contain 0.02% to 0.10% formula (II) and/or formula (III) Eliquis of dimer impurity pass through the present invention step e) to i) method obtain, methods described include will bag The formula of the dimer impurity of formula (IV) and/or formula (V) containing the amount between 2% to 5% determined by HPLC A/A% (VIII) compound is converted into Eliquis.In this case, it is necessary to perform the optional step h) of repetition purification step once Or repeatedly.
The main-process stream of Fig. 1 display synthesis Eliquis, it includes the institute represented in the form of the chemical constitution of impurity and amount Carriage of the impurity Jing Guo the building-up process is stated, the amount is based on art methods.
By the Fig. 2 for the main-process stream for comparing display synthesis Eliquis, the flow includes the method according to the invention (step It is rapid a) to d)), effect caused by the present invention is it is obvious that because in intermediate (IX) and the then impurity in final Eliquis Amount it is more much lower than in existing method (referring to Fig. 1).
It is noted that the amount of the impurity of formula (IV) and formula (V) by methods described seems to increase, particularly by formula (IX) During compound is converted into compound (VIII).Because compared with molecule (VI) and (VII), molecule (IV) and (V) virtue Fragrance increase, so produce increased absorption, i.e. the response increase of detector.Therefore, this increase is only surface, not The truth of the impurity actual amount.
As another aspect of the present invention, it should be taken into account that the dimer impurity of Eliquis includes azo or azoxy Functional group, so as to be azo or azoxy compound, therefore, " the Guideline On The promulgated according to EMEA in 2006 The regulation for the route guidance ICH M7 that Limits Of Genotoxic Impurities " and on June 23rd, 2014 promulgate, very may be used There can be serious carcinogenic risk.
In fact, azoxy compound, which is one group, has formula RN=N+(O-) R common functional group chemical combination Thing, the functional group are present in the dimer impurity of some Eliquis, i.e. formula (VII) dimer impurity, formula (V) dimer Impurity and formula (III) dimer impurity.
Therefore, it is necessary to send the alarm relevant with the potential genetoxic of the dimer impurity.
Experimental section
Compound is named using the primitive rule of IUPAC nomenclatures.
Proton magnetic resonance (NMR) spectrum on Varian instruments 400MHz or on Bruker instruments in 300MHz and 400MHz is recorded.Using residual solvent line as internal standard, chemical shift is reported with ppm (δ).Division peak figure shape is referred to as s, single Peak；D, doublet；T, triplet；Q, quartet；M, multiplet；B, broad peak.Temperature note of the H NMR spectroscopy in the range of 20 DEG C to 30 DEG C Record.
UV-visible (UV-Vis) is obtained by HPLC/DAD detectors.
Following table lists abbreviation used：
T temperature
Eq. equivalent
TEA triethylamines
THF tetrahydrofurans
NMR nuclear magnetic resonance
HPLC high performance liquid chromatographies
LC-MS liquid chromatography-mass spectrometries
UV-Vis UV-visibles
DAD diodes-array detector
Embodiment 1：Prepare formula (IX) compound for including following related dimer impurity：Formula (VI) (E) -1,1'- (4, 4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (morpholinyl -5,6- dihydropyridines -2 (1H) -one) and formula (VII) double (4- ((the 2H)-yl of morpholinyl -2- oxo -5,6- dihydropyridines -1) phenyl) the phenodiazine olefinic oxides of (Z) -1,2-：
Comparing embodiment
Synthesis flow：
Formula (X) compound can be according to Journal Synthetic Communication, volume 2013,43,72- Page 79, in particular according to entitled " 1- (the 4-Aminophenyl) -3- (morpholin-4-yl) -5,6- of page 76 Method disclosed in dihydropyridin-2 (1H)-one (13) " paragraph, prepared since formula (XI) compound.
Therefore, formula (X) compound is prepared according to following methods.
At T=40/45 DEG C, in 20 minutes, by nine hydrated sulfuric acid sodium (197g, 2.5eq.) in water (350mL) Solution is added in the mixture of formula (XI) compound (100g, 1.0eq.) and methanol (1000mL).Stirred at T=40/45 DEG C Mixture is mixed other 2 hours to realize that reaction is completed.Then in decompression and TmaxDistillating mixture at=45 DEG C, until removing 900mL solvents.Gained slurries are cooled to T=20/25 DEG C, is kept stirring for 2 hours in this temperature, then filters, with water (2 × 100mL) wash wet cake.After being dried 10 hours at depressurizing with T=65 DEG C, acquisition 83g formulas (X) compound (92% yield, HPLC A%：Formula (X) compound 96.09%).
The formula (X) compound so obtained contains following dimer impurity：
- 0.48% (HPLC A/A%) formula (VI) dimer impurity,
- 1.39% (HPLC A/A%) formula (VII) dimer impurity.
Formula (X) compound (1.0eq.) described in loading 60g into 4 neck round-bottom flasks (prepares and contains formula as described above (VI) and formula (VII) impurity), be then charged into 51.1g triethylamines (TEA) (2.3 equivalent) and 600mL tetrahydrofurans (THF). At T=0/5 DEG C, solution of the 51g chlorine valeric chloride (1.5 equivalent) in 120mL THF is added within 1-2 hours.At T=0/5 DEG C Other 30 minutes of lower stirring mixture, remain mutually synthermal afterwards, 73.9g potassium tert-butoxides are added within 30-45 minutes The solution of (3.0eq.) in 300mL THF.After being stirred 30 minutes at T=0/5 DEG C, by mixture temperature to after T=20/25 DEG C Maintain again 2 hours.After the completion of examining reaction, in decompression and TmaxBatch of material is distilled at=35 DEG C to remaining 4 volumes (240mL).With 480ml water dilutes gained slurries and remaining 9 volumes (900mL) are arrived in distillation again under the same conditions.
Mixture is stirred at T=20/25 DEG C at least 2 hours, then filter, wet cake is washed with water (2 × 60mL).
After being dried at least 8 hours at depressurizing with T=65 DEG C, 64g formulas (IX) compound (molar yield 82%) is obtained.This Solid contains following dimer impurity：
- 0.17% (HPLC A/A%) formula (VI) dimer impurity,
- 1.9% (HPLC A/A%) formula (VII) dimer impurity.
Embodiment 2：Prepare lower formula (IX) compound, embodiments of the invention
Synthesis flow：
Formula (X) compound can be according to Journal Synthetic Communication, volume 2013,43,72- Page entitled " 1- (the 4-Aminophenyl) -3- (morpholin-4-yl) -5,6- of page 79, particularly the 76th Method disclosed in dihydropyridin-2 (1H)-one (13) " paragraph, prepared since formula (XI) compound.
Therefore, formula (X) compound is prepared according to following methods.
At T=40/45 DEG C, in 45 minutes, by nine hydrated sulfuric acid sodium (197g, 2.5eq.) in water (350mL) Solution is added in the mixture of formula (XI) compound (100g, 1.0eq.) and methanol (1000mL).Stirred at T=40/45 DEG C Mixture is mixed other 2 hours to realize that reaction is completed.Then in decompression and TmaxDistillating mixture at=45 DEG C, until removing 930mL solvents.Gained slurries are cooled to T=20/25 DEG C, is kept stirring for 1 hour in this temperature, then filters, with water (2 × 100mL) wash wet cake.After being dried 10 hours at depressurizing with T=65 DEG C, acquisition 82g formulas (X) compound (91% yield, HPLC A%：Formula (X) compound 96.16%).
The formula (X) compound so obtained contains following dimer impurity：
- 0.66% (HPLC A/A%) formula (VI) dimer impurity,
- 1.50% (HPLC A/A%) formula (VII) dimer impurity.
Formula (X) compound (1.0eq.) described in loading 80g into 4 neck round-bottom flasks (prepares and contains formula as described above (VI) and formula (VII) impurity), be then charged into 68.3g triethylamines (TEA) (2.3 equivalent) and 800mL tetrahydrofurans (THF). At T=0/5 DEG C, solution of the 68.3g chlorine valeric chloride (1.5 equivalent) in 160mL THF is added within 1-2 hours.
Mixture is stirred at T=0/5 DEG C other 30 minutes, remain mutually synthermal afterwards, added within 30-45 minutes Solution of the 98.1g potassium tert-butoxides (3.0eq.) in 400ml mL THF.After being stirred 30 minutes at T=0/5 DEG C, by mixture Maintained other 2 hours after warm to T=20/25 DEG C.After the completion of examining reaction, in decompression and TmaxBatch of material is distilled at=35 DEG C to arrive Remaining 6 volumes (480mL), then under the same conditions again with 560mL ethanolic extractions to remaining 6 volumes (480mL).With 720mL ethanol and 880mL water dilution gained mixture, are then heated to T=40/45 DEG C and in silicon by gained milky white solution Filter in diatomaceous earth, washed (parent material that this solid filter cake is embodiment 3) with the 80mL ethanol preheated.Distillation filter under reduced pressure The solution crossed is to remaining 6 volumes (480mL).Mixture is cooled to T=20/25 DEG C and stirred at least 1 hour in this temperature. Slurries are finally filtered, wet cake is washed with water (2 × 80mL).After being dried at least 8 hours at depressurizing with T=65 DEG C, obtain 90.3g formulas (IX) compound (molar yield 86.8%).
This solid obtained contains following dimer impurity：
-0.04%(HPLC A/A%) formula (VI) dimer impurity,
-0.09%(HPLC A/A%) formula (VII) dimer impurity.
Embodiment 3：Prepare the dimer impurity of lower formula (VI) and lower formula (VII)
The insoluble substance being collected into ethanol solution is enclosed in into the round-bottomed flask equipped with Dean-Stark devices (solid filter cake of embodiment 2, it includes diatomite and the dimer impurity of formula (VI) and formula (VII)) and it is suspended in 600mL In toluene.Backflow is heated the mixture to go to remove water, until reaching solvent boiling point.Mixture is cooled to T=80/90 DEG C simultaneously And filter.Collect solid, it includes the dimer impurity and diatomite of formula (VI) and formula (VII), be fitted into round-bottomed flask and Add 500mL dichloromethane.Heat the mixture to and flow back and filter, washed with dichloromethane (4 × 100mL).Then exist The lower concentrate solution of decompression, obtains residue, and with acetone solution residue to obtain slurries capable of stirring.Filter suspension, 5.8g yellow solids are obtained, after HPLC analyses (Fig. 2), obtain the mixture of following two materials：
- 16.9% (HPLC A/A%) formula (VI) dimer impurity,
- 80.5% (HPLC A/A%) formula (VII) dimer impurity.
Also pass through NMR analysis of mixtures；Because described two impurity differ only in diazo oxidation state difference, because This their 1H chemical shift is overlapping and gained spectrogram shows as the spectrum of single substance.1H-NMR(400MHz,CDCl3, ppm),d:8.32 (dd, J1=8Hz, J2=24Hz 4H), 7.53 (m, 4H), 5.76 (bs, 2H), 3.88 (m, 12H), 2.96 (m,8H),2.57(m,4H)。13C-NMR and DEPT135Aromatic carbon display portion asymmetry on H NMR spectroscopy, and two kinds of materials Aliphatic carbons and carbonyl show identical chemical shift (100MHz, CDCl3,ppm),d:161.4(C),145.7(C),145.1 (C),143.8(C),143.7(C),143.6(C),141.3(C),126.3(CH),125.1(CH),124.6(CH),124.5 (CH),123.3(C),122.8(CH),115.1(CH),114.7(CH),66.8(CH2),50.6(CH2),48.5(CH2),23.4 (CH2).HPLC-MS：11.3min ESI-MS m/z=559 (formula (VII) dimer impurity MW 558 [M+H]+),11.9min ESI-MS m/z=543 (formula (VI) dimer impurity MW542 [M+H]+)。
Embodiment 4：Formula (I) compound is prepared since formula (IX) compound of embodiment 2.The present invention is to Eliquis Effect.
Synthesis flow：
Formula (VIII) compound can according to Journal Synthetic Communication, volume 2013,43, Page entitled " Ethyl1- (4-methoxyphenyl) -7-oxo-6- (4- (2-oxopiperidin- of 72-79 pages, particularly the 78th 1-yl)phenyl)-4,5,6,7-tetr ahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate(2)” Paragraph disclosed in method, prepared since formula (IX) compound prepared according to embodiment 2.
Then, applied according to EP described in EP14189007.9 embodiment 11 or WO2007/001385 embodiment 6 Prepared by method, obtain formula (I) compound Eliquis, it includes the dimer impurity of following Eliquis：
-0.00%Following structure (II) (E) -6,6'- (4,4'- (diazene -1,2- diyls) double (4,1- Asias benzene Base)) double (1- (4- methoxyphenyls) -7- oxo -4,5,6,7- tetrahydrochysene -1H- pyrazolos [3,4-c] pyridine-3-carboxamides)：
-0.01%Following structure (III) double (4- (3- carbamyls -1- (4- the methoxyphenyls) -7- of (Z) -1,2- Oxo -4,5- dihydro-1 h-pyrazoles simultaneously [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
Embodiment 5：Formula (VIII) compound is prepared since formula (IX) compound of embodiment 1.Comparing embodiment.
Synthesis flow：
Formula (VIII) compound can according to Journal Synthetic Communication, volume 1013,43, Page entitled " Ethyl1- (4-methoxyphenyl) -7-oxo-6- (4- (2-oxopiperidin- of 71-79 pages, particularly the 78th 1-yl)phenyl)-4,5,6,7-tetr ahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate(2)” Paragraph disclosed in method, prepared since formula (IX) compound prepared according to embodiment 1.
Formula (VIII) compound of acquisition contains following dimer impurity：
(E) -6,6'- (4,4'- (diazene -1,2- diyls) double (4,1- Asias benzene of -0.59% following structure (IV) Base)) double (1- (4- methoxyphenyls) -7- oxo -4,5,6,7- tetrahydrochysene -1H- pyrazolo [3,4-c] Nicotinicum Acidums ethyl esters)：
(Z) -1,2- of -1.61% following structure (V) it is double (4- (3- (ethoxy carbonyl) -1- (4- methoxyphenyls) - 7- oxo -4,5- dihydro-1 h-pyrazoles simultaneously [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
Embodiment 6：The purifying of formula (VIII) compound.The effect of the present invention.
Load 10g 1- (4- methoxyphenyls) -7- oxos -6- (4- (2- oxo-piperidine -1- bases) into 4 neck round-bottom flasks Phenyl) -4,5,6,7- tetrahydrochysene -1H- pyrazolos [3,4-c] Nicotinicum Acidum ethyl esters (0.02mol) (formula (VIII) compound), It contains the dimer of 0.21% (HPLC A/A%) formula (IV) and 0.10% (HPLC A/A%) formula (V) dimer impurity.
The compound is suspended in 100mL ethanol and heats the mixture to backflow, produces milky white solution.Add Add activated carbon (1g), and after flowing back other 30 minutes, filter mixture while hot, washed with the ethanol (2 × 20mL) of preheating.Will Filtered solution is cooled to T=20/25 DEG C and stirred at least 2 hours in this temperature.Then slurries are filtered, with ethanol (2 × 10mL) wash filter cake.Wet solid is dried under vacuum at T=65 DEG C.(70% mole of 7.0g formulas (VIII) compound of gained Yield) contain：
-0.06%(4,4'- (diazene -1,2- diyls) is double, and (4,1- is sub- by (E) -6,6'- of (HPLC A/A%) formula (IV) Phenyl)) double (1- (4- methoxyphenyls) -7- oxo -4,5,6,7- tetrahydrochysene -1H- pyrazolos [3,4-c] Nicotinicum Acidum second Ester)
-0.03%Double (4- (3- (ethoxy carbonyl) -1- (the 4- methoxybenzenes of (Z) -1,2- of (HPLC A/A%) formula (V) Base) -7- oxo -4,5- dihydro-1 h-pyrazoles simultaneously [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide.
Embodiment 7：The purifying of formula (VIII) compound.The effect of the present invention.
From miscellaneous containing 0.59% (HPLC A/A%) formula (IV) dimer and 1.80% (HPLC A/A%) formula (V) dimer Formula (VIII) compound of matter starts to repeat the embodiment.
Gained formula (VIII) compound contains：
-0.11%(HPLC A/A%) formula (IV) dimer impurity
-0.36%(HPLC A/A%) formula (V) dimer impurity.
Can program is one or many to be further purified the compound (VIII) by repeating described in embodiment 6.
Embodiment 8：
Determine the analysis method of the amount of following material：
The dimer impurity of formula (VI) and formula (VII) in-formula (IX) and the compound of (X),
The dimer impurity of formula (IV) and formula (V) in-formula (VIII) compound,
The dimer impurity of formula (II) and formula (III) in-formula (I) Eliquis.
The compound can be identified and monitored by following HPLC methods：
Chromatographic condition：
Using above-mentioned condition, it is contemplated that retention time it is as follows：
Compound Retention time Relative retention time
Formula (IX) compound 8.02min 1.14
Formula (X) compound 4.70min 0.67
Formula (XI) compound 6.30min 0.90
Formula (VI) dimer impurity 11.67min 1.66
Formula (VII) dimer impurity 11.10min 1.58
Formula (VIII) compound 11.37min 1.62
Formula (IV) dimer impurity 18.31min 2.60
Formula (V) dimer impurity 17.75min 2.52
Formula (I) Eliquis 7.03min 1.00
Formula (II) dimer impurity 12.31min 1.75
Formula (III) dimer impurity 12.00min 1.71
The dimer impurity described in description and embodiments on the compound of formula (I), (VIII), (IX) and (X) Amount, the amount particularly represented with HPLC A/A%, can according to described in the present embodiment 8 HPLC methods carry out.
For the LC-MS methods for the dimer impurity for determining Eliquis.
LC parameters
Embodiment 9：Formula (IV) two is prepared since the mixture of formula (VI) dimer impurity and formula (VII) dimer impurity Aggressiveness impurity and formula (V) dimer impurity.
Synthesis flow：
Added into round-bottomed flask：Pass through formula (VI) dimer impurity and formula for reforming embodiment 2 and 3 to prepare on a large scale (VII) dimer impurity mixture (14g, 25mmol, 1.0eq., consider formula (VII) dimer impurity material molecular weight), Formula (XII) compound (90g, 350mmol, 7.0eq.), triethylamine (76g, 750mmol, 15.0eq.), KI (8.5g, 51mmol, 1.0eq.) and ethyl acetate (1960ml).Suspension is heated to flow back and stirred 16 hours, is consequently cooled to T =20/25 DEG C and filter to remove insoluble substance, wash filter cake with ethyl acetate (3 × 20ml).
Be slowly added at T=20/25 DEG C into resulting solution the 32%HCl aqueous solution (72ml, d=1.16g/ml) and Mixture is stirred in this temperature 30 minutes.
Filter slurries and collected wet solid is suspended in saturated solution of sodium carbonate (50ml) at T=20/25 DEG C In.After this temperature stirs 30min, filter mixture and sequentially washed with saturated solution of sodium carbonate (50ml) and water (20ml) Wet cake.
After being dried 8 hours at depressurizing with T=65 DEG C, 8.4g dimer impurities mixture (40% yield, HPLC are obtained A%：The dimer (MW 824) 71.3% of formula (V)；Dimer (MW 808) the 12.2%HPLC A/A% of formula (IV)).
Embodiment 10：Formula (V) dimer impurity is prepared since formula (III) dimer impurity and formula (II) dimer impurity With formula (IV) dimer impurity.
Synthesis flow：
Into 1L stainless steel autoclaves add from carry out on a large scale embodiment 9 acquisition formula (IV) dimer impurity with The mixture of formula (V) dimer impurity (15g, 18.5mmol, 1.0eq., considers the dimer impurity materials of MW 824 of formula (V) Molecular weight) and propane diols (600ml).After nitrogen purging, at T=20/25 DEG C, add in ammonia to p=1.5/2.0 bars of stabilization Portion's pressure.Then T=120 DEG C is heated the mixture to, internal pressure is reached P=9 bars.
After this temperature 40 hours, mixture is cooled to T=20/25 DEG C and with propane diols (30ml), water (360ml) Diluted with ethanol (90ml).Slurries 1 hour obtained by stirring, finally filters, filter cake is washed with water (60mL) at T=20/25 DEG C. Wet stock is dried in the case where depressurizing with T=65 DEG C 8 hours, obtains 10.3g dimer impurities mixture (74% yield, HPLC A%： The dimer MW 766 48.53% of formula (III)；The dimer MW 750 34.6% of formula (II)).
Embodiment 11：The chromatographic isolation of dimer impurity mixture and the analysis test of single contaminant.
Separate the analysis method of following dimer impurity and the sign of each single dimer impurity：
1. the mixture of formula (VI) dimer impurity and formula (VII) dimer impurity,
2. the mixture of formula (IV) dimer impurity and formula (V) dimer impurity
3. the mixture of formula (II) dimer impurity and formula (III) dimer impurity.
1. the chromatographic isolation of formula (VI) dimer impurity and the mixture of formula (VII) dimer impurity
The separation of the mixture obtained in embodiment 3 passes through preparation HPLC (Agilent Technologies 1200 series) realized using following condition：
Chromatographic condition：
The sign of formula (VII) dimer impurity
Retention time：11.8min (is obtained) by means of above-mentioned chromatographic condition.
LC-MS(ESI+)：M/z=559, [M+H]+
1H NMR(400MHz,CDCl3)δ(ppm):8.34 (d, J=8.8Hz, 2H)；8.27 (d, J=8.8Hz, 2H)； 7.51(m,4H)；5.73(m,2H)；3.85(m,12H)；2.93(m,8H)；2.56(m,4H).
13C and DEPT135NMR(100MHz,CDCl3)δ(ppm):161.3(C)；145.6(C)；145.05(C)；143.7 (C)；143.6(C)；143.5(C)；141.3(C)；126.3(CH)；124.6(CH)；124.5(CH)；122.7(CH)；115.2 (CH)；114.9(CH)；66.7(CH2)；50.5(CH2)；48.4(CH2)；23.4(CH2)。
UV-Vis spectrum：
There is obtained the maximum absorption under 350-360nm.
The sign of formula (VI) dimer impurity
Retention time：14.3min (is obtained) by means of above-mentioned chromatographic condition.
LC-MS(ESI+)：M/z=543, [M+H]+
1H NMR(400MHz,CDCl3)δ(ppm):8.00(m,4H)；7.56(m,4H)；5.37(m,2H)；3.73(m, 12H)；3.27(m,8H)；2.96(m,4H).
2. the chromatographic isolation of formula (IV) dimer impurity and the mixture of formula (V) dimer impurity
The separation of the mixture obtained in embodiment 9 passes through preparation HPLC (Agilent Technologies 1200 series) realized using following condition：
Chromatographic condition：
The sign of formula (V) dimer impurity
Retention time：3.0min (is obtained) by means of above-mentioned chromatographic condition.
LC-MS(ESI+)：M/z=825, [M+H]+
1H NMR(400MHz,CDCl3)δ(ppm):8.32 (d, J=9.2Hz, 2H)；8.24 (d, J=9.2Hz, 2H)； 7.48(m,8H)；6.95 (dd, J1=8.8Hz J2=2Hz, 4H)；4.49 (q, J=6.8Hz, 4H)；4.21(m,4H)；3.84 (s,6H)；3.36(m,4H)；1.46 (t, J=6.8Hz, 6H).
13C and DEPT135NMR(100MHz,CDCl3)δ(ppm):162.1(C)；162.0(C)；160.0(C)；159.9 (C)；157.1(C)；145.5(C)；144.7(C)；142.5(C)；141.7(C)；139.1(C)；139.0(C)；126.94 (CH)；126.93(CH)；126.4(CH)；125.2(CH)；125.1(CH)；122.79(CH)；113.73(CH)；113.71 (CH)；61.33(CH2)；61.29(CH2)；55.5(CH3)；50.9(CH2)；21.6(CH2)；14.4(CH3)；
UV-Vis spectrum：
There is obtained the maximum absorption under 356-360nm.
The sign of formula (IV) dimer impurity
Retention time：4.0min (is obtained) by means of above-mentioned chromatographic condition.
LC-MS(ESI+)：M/z=809, [M+H]+
1H NMR(400MHz,CDCl3)δ(ppm):7.94 (d, J=8.8Hz, 4H)；7.50(m,8H)；6.95 (d, J= 9.2Hz,4H)；4.49 (q, J=7.2Hz, 4H)；4.22(m,4H)；3.84(s,6H)；3.36(m,4H)；1.46 (t, J= 7.2Hz,6H)。
UV-Vis spectrum：
There is obtained the maximum absorption under 356-360nm.
3. the chromatographic isolation of formula (II) dimer impurity and the mixture of formula (III) dimer impurity
The separation of the mixture obtained in embodiment 10 passes through preparation HPLC (Agilent Technologies 1200 series) realized using following condition：
Chromatographic condition：
Because the low-solubility of these materials, prepare 5mg/mL solution and filter insoluble substance with obtain clarification it is molten Liquid.
The sign of formula (III) dimer impurity
Retention time：13.6min (is obtained) by means of above-mentioned chromatographic condition.
LC-MS(ESI+)：M/z=767, [M+H]+
1H NMR(400MHz,CDCl3)δ(ppm):8.28 (d, J=9.2Hz, 2H)；8.18 (d, J=8.8Hz, 2H)； 7.76(bs,2H)；7.63-7.52(m,8H)；7.47(bs,2H)；7.02 (d, J=8.8Hz, 4H)；4.17(m,4H)；3.82 (s,6H)；3.25(m,4H).
UV-Vis spectrum：Shown in Fig. 4
There is obtained the maximum absorption under 360-370nm.
The sign of formula (II) dimer impurity
Retention time：14.6min (is obtained) by means of above-mentioned chromatographic condition.
LC-MS(ESI+)：M/z=751, [M+H]+
1H NMR(400MHz,CDCl3)δ(ppm):7.93 (d, J=8.8Hz, 4H)；7.76(bs,2H)；7.63-7.52 (m,8H)；7.47(bs,2H)；7.02 (d, J=8.8Hz, 4H)；4.17(m,4H)；3.82(s,6H)；3.25(m,4H).
UV-Vis spectrum：Shown in Fig. 3
There is obtained the maximum absorption under 350-360nm.
MS and UV the spectrum measure of dimer impurity
LC parameters
In embodiment 1, there is a large amount of formulas (VI) and formula using the art methods for preparing formula (IX) compound, acquisition (VII) compound of dimer impurity.
In example 2, using the teachings of the present invention, i.e. solution of the filtration compounds (IX) in second alcohol and water, gained The content of each dimer impurity of formula (VI) and formula (VII) is less than 0.10% in formula (IX) compound.
Therefore, can be by comparing embodiment 1 and embodiment 2, protrusion is related to step a) to effect of the invention d).
In embodiment 3, the impurity of formula (VI) and formula (VII) is by removing in embodiment 2 two by formula (VI) and (VII) It is prepared by solid that the mixture of aggressiveness impurity is formed.
Embodiment 4 provides evidence and proves to use the inventive method, formula (II) that final product Eliquis includes and (III) content of dimer impurity is less than 0.10%.
Embodiment 5 describes to prepare formula (VIII) compound according to art methods, and obtains to include and largely (be higher than 0.50%, HPLC A/A%) formula (IV) and formula (V) dimer impurity compound (VIII).
Embodiment 6 provides the evidence for proving effect of the present invention, because according to the step e) of the inventive method to i) purifyingization Compound (VIII), so as to which the primary quantity of impurity (IV) and (V) is reduced into 0.06% He respectively from 0.21% and 0.10% 0.03%.
Embodiment 7 provides the evidence for proving effect of the present invention again, because according to the inventive method purifying compound (VIII), so as to distinguish impurity (IV) and (V) primary quantity from 0.59% and 1.80% in the compound (VIII) of separation It is reduced to 0.11% and 0.36%.It should be evident that the product (VIII) of this experiment can be by carrying out such as the He of embodiment 6 to it Identical experiment described in optional step h) is one or many to be further purified.
By the result analyzed above-mentioned experiment He provided, the water of dimer impurity in Eliquis is especially considering that It is flat, it will be appreciated that the inventive method provided removing dimer impurity so as to providing the Ah substantially free of dimer impurity Technique effect in piperazine sand class.

Claims (24)

1. a kind of method for preparing formula (I) Eliquis,
The Eliquis includes the dimer impurity of the following Eliquis less than 0.10%：
(E) -6,6'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (1- (4- methoxies of following structure (II) Base phenyl) -7- oxos -4,5,6,7- tetrahydrochysene -1H- pyrazolos [3,4-c] pyridine-3-carboxamide)：
And/or the dimer impurity of following Eliquis：
(Z) -1,2- of following structure (III) it is double (4- (3- carbamyls -1- (4- methoxyphenyls) -7- oxo -4,5- dihydros - 1H- pyrazolos [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
Wherein it the described method comprises the following steps：
A) morpholinyl -1- (4- (2- oxo-piperidine -1- bases) phenyl) -5,6- dihydropyridines -2 (1H) -one of formula (IX) is prepared Solution in the mixture of C1-C3 alcohol and water：
B) the middle solution prepared of the step a) is filtered to remove following dimer impurity：
(E) -1,1'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (morpholinyl -5,6- two of formula (VI) Pyridinium hydroxide -2 (1H) -one) and/or formula (VII) the double (4- (morpholinyl -2- oxos -5,6- two of dimer impurity (Z) -1,2- Pyridinium hydroxide -1 (2H)-yl) phenyl) phenodiazine olefinic oxide：
C) solution of formula (IX) compound comprising purifying is obtained in filtrate,
D) the step c) formula (IX) compound is converted into formula (I) Eliquis；
Or alternatively, the step a) is replaced to d) with following steps：
E) 1- (4- the methoxyphenyls) -7- oxos -6- (4- (2- oxo-piperidine -1- bases) phenyl) -4,5 of formula (VIII) is prepared, Solution of 6,7- tetrahydrochysene -1H- pyrazolo [3,4-c] the Nicotinicum Acidum ethyl esters in C1-C4 alcohol：
F) the middle solution prepared of the step e) is filtered to remove following dimer impurity：
(E) -6,6'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (1- (4- methoxybenzenes of lower formula (IV) Base) -7- oxo -4,5,6,7- tetrahydrochysene -1H- pyrazolo [3,4-c] Nicotinicum Acidums ethyl ester) and/or the dimer of formula (V) it is miscellaneous Matter (Z) -1,2- it is double (4- (and 3- (ethoxy carbonyl) -1- (4- methoxyphenyls) -7- oxo -4,5- dihydro-1 h-pyrazoles simultaneously [3, 4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
G) solution of formula (VIII) compound comprising purifying is obtained in filtrate,
H) optionally, formula (VIII) compound of the purifying of solid form, and repeating said steps e) to g) one are separated It is secondary or multiple,
I) formula (VIII) compound by step g) or h) is converted into formula (I) Eliquis.
2. according to the method for claim 1, wherein in the step a), with the formula (IX) compound phase ratio, C1-C3 The amount of alcohol is 5 to 20 volumes.
3. method according to any one of claim 1 to 2, wherein in the step a), with the formula (IX) chemical combination Thing is compared, and the amount of water is 1 to 15 volume.
4. according to the method in any one of claims 1 to 3, wherein in the step a), the amount of C1-C3 alcohol for 5 to 20 volumes, and the amount of water is 1 to 15 volume, both of which and the formula (IX) compound phase ratio.
5. method according to any one of claim 1 to 4, wherein in the step a), the C1-C3 alcohol is second Alcohol.
6. method according to any one of claim 1 to 5, wherein the step b) enters in 30 DEG C to 50 DEG C of temperature OK.
7. according to the method for claim 1, wherein in the step e), the C1-C4 alcohol is ethanol.
8. the method according to claim 1 or 7, wherein the step f) enters in the temperature of 40 DEG C to the alcohol of boiling point OK.
9. method according to any one of claim 1 to 8, wherein in the step a) or described steps e), enter one Step addition filter aid and/or activated carbon.
10. method according to any one of claim 1 to 9, wherein the step b) or step f) are by means of diatomite Plate is carried out.
11. method according to any one of claim 1 to 10, wherein Eliquis include less than 0.10% following Ah The dimer impurity of piperazine sand class：Double (4- (3- carbamyls -1- (4- the methoxyphenyls) -7- oxygen of (Z) -1,2- of lower formula (III) Generation -4,5- dihydro-1 h-pyrazoles simultaneously [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
The precursor dimer impurity of removal formula (VII) wherein in the step b), or the removal formula (V) in the step f) Precursor impurity.
12. the method according to any one of claim 1 to 11, wherein Eliquis include the formula (II) less than 0.01% And/or the dimer impurity of formula (III).
Determined 13. the method according to any one of claim 1 to 11, wherein Eliquis contain by HPLC A/A% 0.00% to 0.02% formula (II) and/or formula (III) dimer impurity, methods described includes the formula (IX) chemical combination Thing is converted into Eliquis, and wherein described formula (IX) compound include by HPLC A/A% determine 0.1% to 2% Between the formula (VI) of amount and/or the dimer impurity of formula (VII).
Determined 14. the method according to any one of claim 1 to 11, wherein Eliquis contain by HPLC A/A% 0.02% to 0.10% formula (II) and/or formula (III) dimer impurity, methods described includes the formula (IX) chemical combination Thing is converted into Eliquis, and wherein described formula (IX) compound include by HPLC A/A% determine 2% to 5% it Between the formula (VI) of amount and/or the dimer impurity of formula (VII).
15. the method according to any one of claim 1 to 14, wherein being obtained in the step d) or described steps i) Eliquis there is N-1 forms.
16. the dimer impurity of Eliquis or its precursor, it is selected from：
A (E) -6,6'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (1- (4- methoxyl groups of formula (II) under) Phenyl) -7- oxos -4,5,6,7- tetrahydrochysene -1H- pyrazolos [3,4-c] pyridine-3-carboxamide)：
B under) (Z) -1,2- of formula (III) it is double (4- (3- carbamyls -1- (4- methoxyphenyls) -7- oxo -4,5- dihydros - 1H- pyrazolos [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
C under) (E) -1,1'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) of formula (VI) it is double (morpholinyl -5, 6- dihydropyridines -2 (1H) -one)：
D double (4- ((the 2H)-yl of morpholinyl -2- oxo -5,6- dihydropyridines -1) phenyl) two of (Z) -1,2- of formula (VII) under) Diazenium oxide：
E (E) -6,6'- (4,4'- (diazene -1,2- diyls) is double (4,1- phenylenes)) double (1- (4- methoxyl groups of formula (IV) under) Phenyl) -7- oxo -4,5,6,7- tetrahydrochysene -1H- pyrazolo [3,4-c] Nicotinicum Acidums ethyl ester)：
F under) (Z) -1,2- of formula (V) it is double (4- (3- (ethoxy carbonyl) -1- (4- methoxyphenyls) -7- oxo -4,5- dihydros - 1H- pyrazolos [3,4-c] pyridine -6 (7H)-yl) phenyl) phenodiazine olefinic oxide：
17. the dimer impurity of dimer impurity according to claim 16, wherein Eliquis or its precursor is yellow Powder.
18. the dimer impurity according to any one of claim 16 to 17, it is (Z) -1,2- of the lower formula (III) It is double (4- (3- carbamyls -1- (4- methoxyphenyls) -7- oxo -4,5- dihydro-1 h-pyrazoles simultaneously [3,4-c] pyridine -6 (7H) - Base) phenyl) phenodiazine olefinic oxide：
19. a kind of dimer impurity for being used to determine the Eliquis of lower formula (II) and/or lower formula (III) in Eliquis Method：
It comprises the following steps：
A) formula (II) of known quantity and/or the dimer impurity of formula (III) are added into Eliquis sample,
B) HPLC or LC/MS analyses are carried out to the Eliquis sample of the step a),
C) HPLC the or LC/MS peaks of dimer impurity are detected；
Or,
A1 dimer impurity) is analyzed by means of HPLC or LC/MS,
B1 the Eliquis sample) is analyzed by means of HPLC or LC/MS,
C1 HPLC the or LC/MS peaks of dimer impurity) are detected by comparing retention time or relative retention time；
Or,
Detection [M+1] is analyzed by means of LC/MS+Equal to 751 or 767amu peak.
20. a kind of dimer impurity of the Eliquis of lower formula (II) being used in quantitative Eliquis and/or lower formula (III) Method：
It comprises the following steps：
A) by means of corresponding to each single the two of formula (II) and/or formula (III) in HPLC or LC/MS measurement Eliquis samples The peak area of aggressiveness impurity, the Eliquis sample have these compounds of unknown quantity,
B) by means of HPLC or LC/MS measurements corresponding to the dimer impurity of the formula (II) containing known quantity and/or formula (III) The peak area of " normative reference thing ",
C) Ah is determined with the middle areas measured of the step b) by the area of measurement in the step a) The amount of dimer impurity in piperazine sand class.
21. the method according to any one of claim 19 or 20, wherein the dimer impurity of the Eliquis is under Formula (III) represents：
22. the dimer impurity of Eliquis below is used as being used to identify and/or quantifying described two in formula (I) Eliquis The purposes of " the reference mark thing " or " normative reference thing " of aggressiveness impurity,
A) dimer impurity of formula (II)：
B) dimer impurity of formula (III)：
23. purposes according to claim 20, wherein the dimer impurity of the Eliquis is the formula (III) dimerization Body impurity：
24. a kind of dimer impurity of the Eliquis of lower formula (II) being used in quantitative Eliquis and/or lower formula (III) The method of amount between 1 to 100 PPM：
Wherein described measure is carried out by LC/MS.
CN201680026473.6A 2015-07-20 2016-07-18 The dimer impurity and its minimizing technology of Eliquis Pending CN107567447A (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP15177517.8 2015-07-20
EP15177517.8A EP3064497A1 (en) 2015-07-20 2015-07-20 Dimer impurities of apixaban and method to remove them
PCT/EP2016/067083 WO2017013079A1 (en) 2015-07-20 2016-07-18 Dimer impurities of apixaban and method to remove them

Publications (1)

Publication Number Publication Date
CN107567447A true CN107567447A (en) 2018-01-09

Family

ID=53758032

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201680026473.6A Pending CN107567447A (en) 2015-07-20 2016-07-18 The dimer impurity and its minimizing technology of Eliquis

Country Status (6)

Country Link
US (2) US10385049B2 (en)
EP (2) EP3064497A1 (en)
JP (1) JP6692408B2 (en)
CN (1) CN107567447A (en)
ES (1) ES2718038T3 (en)
WO (1) WO2017013079A1 (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3064497A1 (en) * 2015-07-20 2016-09-07 F.I.S.- Fabbrica Italiana Sintetici S.p.A. Dimer impurities of apixaban and method to remove them
CN108645933B (en) * 2018-05-24 2020-10-16 河北常山生化药业股份有限公司 Gas chromatography detection method for residual solvent in apixaban bulk drug
CN114910596B (en) * 2021-02-07 2024-01-26 南京正大天晴制药有限公司 Analysis method for determining related substances of pyridone compounds

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003049681A2 (en) * 2001-12-10 2003-06-19 Bristol-Myers Squibb Company Synthesis of 4,5-dihydro-pyrazolo [3,4-c] pyrid-2-ones
CN103626759A (en) * 2012-08-24 2014-03-12 上海医药工业研究院 Preparation method of apixaban intermediate
WO2014108919A2 (en) * 2013-01-09 2014-07-17 Msn Laboratories Limited NOVEL INTERMEDIATE AND POLYMORPHS OF 1-(4-METHOXYPHENYL)-7-OXO-6-[4-(2-OXOPIPERIDIN-1-YL)PHENYL]-4,5,6,7-TETRAHYDRO-1H-PYRAZOLO[3,4-c] PYRIDINE-3-CARBOXAMIDE AND PROCESS THEREOF
CN104030972A (en) * 2014-06-17 2014-09-10 河北序能生物技术有限公司 Synthesis method for pyridone

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7396932B2 (en) 2004-09-28 2008-07-08 Bristol-Myers Squibb Company Process for preparing 4,5-dihydro-pyrazolo[3,4-c]pyrid-2-ones
CN101967145B (en) * 2010-09-09 2012-07-04 华东理工大学 Method for preparing antithrombotic medicament apixaban
CN103626689B (en) * 2012-08-24 2016-03-09 上海医药工业研究院 A kind of preparation method of intermediate of Eliquis
CN104109165A (en) * 2013-04-19 2014-10-22 四川海思科制药有限公司 4,5-dihydro-pyrazolo[3,4-c]pyrid-2-one derivatives, and preparation method and application thereof
CN104341336B (en) * 2013-08-02 2018-10-16 上海科胜药物研发有限公司 A kind of new method of synthesis Eliquis important intermediate
CN104650072B (en) * 2013-11-18 2016-03-16 成都苑东生物制药股份有限公司 A kind of pyridine derivatives
CN103739541B (en) * 2014-01-07 2016-01-20 上海现代制药股份有限公司 The preparation method of 5,6-dihydro-3-(4-morpholinyl)-1-[4-(2-oxo-piperidino) phenyl]-2 (1H)-pyridones
CN104045637B (en) * 2014-04-18 2016-04-06 河北科技大学 A kind of preparation method of Eliquis
EP3064497A1 (en) * 2015-07-20 2016-09-07 F.I.S.- Fabbrica Italiana Sintetici S.p.A. Dimer impurities of apixaban and method to remove them

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003049681A2 (en) * 2001-12-10 2003-06-19 Bristol-Myers Squibb Company Synthesis of 4,5-dihydro-pyrazolo [3,4-c] pyrid-2-ones
CN103626759A (en) * 2012-08-24 2014-03-12 上海医药工业研究院 Preparation method of apixaban intermediate
WO2014108919A2 (en) * 2013-01-09 2014-07-17 Msn Laboratories Limited NOVEL INTERMEDIATE AND POLYMORPHS OF 1-(4-METHOXYPHENYL)-7-OXO-6-[4-(2-OXOPIPERIDIN-1-YL)PHENYL]-4,5,6,7-TETRAHYDRO-1H-PYRAZOLO[3,4-c] PYRIDINE-3-CARBOXAMIDE AND PROCESS THEREOF
CN104030972A (en) * 2014-06-17 2014-09-10 河北序能生物技术有限公司 Synthesis method for pyridone

Also Published As

Publication number Publication date
EP3280714B1 (en) 2019-02-06
EP3280714A1 (en) 2018-02-14
US10590133B2 (en) 2020-03-17
ES2718038T3 (en) 2019-06-27
US20180118737A1 (en) 2018-05-03
JP6692408B2 (en) 2020-05-13
US10385049B2 (en) 2019-08-20
EP3064497A1 (en) 2016-09-07
WO2017013079A1 (en) 2017-01-26
JP2018524385A (en) 2018-08-30
US20190367509A1 (en) 2019-12-05

Similar Documents

Publication Publication Date Title
CA3039090C (en) Process for preparing btk inhibitors
CN102971325A (en) Compounds for the reduction of beta-amyloid production
EP3271362B1 (en) Heterocyclic compounds useful as inhibitors of tnf
CN107567447A (en) The dimer impurity and its minimizing technology of Eliquis
US20100010215A1 (en) Introverted CB[N] Compounds
EP2720989B1 (en) Deuterated 1-piperazino-3-phenyl indanes for treatment of schizophrenia
Garrier et al. First enantiopure calix [6] aza-cryptand: synthesis and chiral recognition properties towards neutral molecules
Mobian et al. Towards the stereoselective synthesis of inherently chiral pseudorotaxanes
CN113614090A (en) Rapamycin derivatives containing isothiazolidine 1, 1-dioxide and 1, 4-butane sultone and uses thereof
Caccamese et al. Inherently chiral α-picolyloxy-p-tert-butylcalix [5] arene crown ethers: Synthesis, structure proof, and enantioselective HPLC resolution
Elkamhawy et al. Pyrazinyl ureas revisited: 1-(3-(Benzyloxy) pyrazin-2-yl)-3-(3, 4-dichlorophenyl) urea, a new blocker of Aβ-induced mPTP opening for Alzheimer's disease
Glenny et al. Improved synthetic methods to mixed-donor thiacrown ethers
CN109053724A (en) The purification process and purposes of the preparation method and Tadalafei of a kind of impurity prepared during Tadalafei and the impurity
CN105849106A (en) Key intermediates and impurities of the synthesis of apixaban: apixaban glycol esters
Németh et al. Synthesis and pKa determination of new enantiopure dimethyl‐substituted acridino‐crown ethers containing a carboxyl group: Useful candidates for enantiomeric recognition studies
Alešković et al. Synthesis, structural characterization, and anion binding ability of sterically congested adamantane-calix [4] pyrroles and adamantane-calixphyrins
CN114773348A (en) Preparation method and intermediate of midazolam
Dolenský et al. Synthesis and characterisation of a new naphthalene tris-Tröger’s base derivative—a chiral molecular clip
Kanger et al. Asymmetric synthesis of novel C2-symmetric bimorpholines
EP4360660A1 (en) Process for the preparation of a gadolinium contrast agent
CN107488144B (en) Molecule capable of specifically binding and inhibiting Tau protein aggregation and preparation method and application thereof
Bedürftig et al. Synthesis and receptor binding studies of 3-substituted piperazine derivatives
Tuchalski et al. X-ray investigations of nebivolol and its isomers
Argese et al. Stereochemistry of the intermediates in the synthesis of 1, 4, 7, 10-tetraazacyclododecane from triethylenetetramine, glyoxal and diethyl oxalate
CN103848847B (en) A kind of method that improvement prepares Olanzapine and its crystal formation II

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication

Application publication date: 20180109

RJ01 Rejection of invention patent application after publication